Mycotoxins: Zearalenone
*This article is not medical advice. Before starting on any health related regimen, seek the advice of your Primary Care Physician or an M.D.
Zearalenone
The mycotoxin zearalenone (ZON) is produced by Fusarium mold species. I chose to highlight this mycotoxin following my blog articles on Ochratoxin A (OTA) for three reasons. First, I often see ZON elevated along with OTA on urine-based mycotoxin tests. Second, when it comes to food contamination, ZON and OTA are found on the same grains. Third, it’s effects on the body are significant, given that it inhibits copper transporters, depletes vitamin B2, causes blood glucose dysregulation, and blocks a key gene (GPX4) which can lead to high mitochondrial oxidative stress and Ferroptosis.
“The results show that Zearaleone induced an increase in ROS generation and lipid peroxidation, and a decrease in levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione (GSH). Zearalenone also causes B2 deficiency and inhibits copper transport”
Why read this article?
You may find this article valuable if you:
Want to learn more about how Zearalenone effects the body
Want to learn about detoxification of Zearalenone
Want to learn what binders can be used to bind Zearelanone
Highlights About Zearalenone:
Causes ferroptosis (un-mitigated cell death leading to large weight loss) by inhibiting gpx4...."ZEA also downregulated the expression of Nrf2, SLC7A11, and GPX4, and .... resulting in the accumulation of lipid peroxides" [15].....
Inhibits glutamate transport and causes NMDA activation, via reduced SLC7A11 transport
Causes copper transport dysregulation [8]
Zearalenone is a xenoestrogen that can contribute to hyperestrogenemia. High estrogen levels can significantly impact the Bile Salt Export Pump (BSEP), which can contribute to Herxheimer reactions and other health consequences of poor bile salt excretion and impaired liver detoxification
Activates the inflammasome (NLRP3) and is responsible for the maturation and secretion of the pro-inflammatory cytokines IL-1β and IL-18, which play a role in IBD. [19]
Activates cytokines (i.e. high IL,4, IL-10, TNFA, INFG; but not IL-8) [15] Elevated IL-10 also disrupts heme synthesis.
Contributes to insulin resistance / blood glucose issues. ZON blocks Glut4 (needed for glucose uptake), leading to dysregulated blood glucose. [23]
Leads to metabolic reprogramming as well as an increase in lactic acid production and NADPH oxidase (NADPH Oxidase= NOX = Super Oxide, and consumes NAD) activity
High lactic acid decreases the activity of proline oxidase, which catalyzes proline, an amino acid that participates in the biochemical transformation of arginine, a source of nitric oxide (NO). NO plays an important function in the regulation of the intestinal barrier function, and decreased proline metabolism can compromise intestinal integrity and lead to intestinal damage
Depletes vitamin B2. Metabolic reprogramming contributes to alterations in the Krebs cycle, leading to accumulation of succinic acid (requires B2 to be cleared), a decrease in the concentrations of reduced forms of B2/flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NAD), and uncoupling of the respiratory chain. Reactive oxygen species are most dynamically formed in complexes I and III of the electron transport chain
Short-term ZON exposure in mice caused intestinal flora imbalance and altered intestinal mucosal immune function, inducing a severe mucosal inflammatory response
ZON has been shown to increase oxidative stress and reduce the activity of important anti oxidant enzymes such as GPX (1-8) and SOD (1-3)
Zearalenone can activate the pregnane X receptor, thereby increasing the transcription of many genes, including Cytochrome P 450 enzymes. Zearalenone has been found to affect hundreds of genes with a mix up-regulation and down-regulation of cellular signaling pathways, cytokine networks, and the inflammation response.
Organ Damage Due To Zearalenone
Studies have shown that zearalenone damages the reproductive organs, intestines, immune system, liver, kidney and spleen. In animals, it can induce an enlarged uterus, ovarian atrophy, endometrial changes, vaginal prolapse, swelling of the vulva and vagina , enlarged breast glands, anestrus , decreased fertility, increased embryonic and fetal death, and abortion.
Food and Cooking Considerations for Zearalenone :
Fusarium mold growth on soy can lead to contamination with ZON, possibly increasing the estrogenic potential. Corn and wheat are also often contaminated with Zearalenone. Note that high relative humidity during storage favors production of Zearalenone.
Zearalenone is relatively resistant to degradation, unfortunately, and it is known for being fairly stable under normal cooking temperatures. ZON is susceptible to alkaline conditions, or to being heated under a high degree of pressure. Thus, high temperatures, milling, dehulling, fermentation, and UV light all help lower the Zearalenone content.
Binders for Zearalenone
Cholestyramine / Chitosan / Welchol
Saccharomyces cerevisiae
Saccharomyces cerevisiae cell walls are used to bind aflatoxins, ochratoxin, and zearalenone in animal foods. (it also acts as a competitor for C. diff and Candida).
The alpha mannan oligosaccharides and Beta-D-glucan in the cell wall help bind Zearalenone. More acidic conditions favor mycotoxin adsorption on yeast cell walls in simulated gastrointestinal environments.
Saccharomyces boulardii can also help biotransform zearalanone and improve clearance this way, although the S. cerevisiae strains CS, LL74 and LL83 were found to most effective for this
Gluconamannan has been shown to be somewhat helpful (given the effect on Beta-D-Glucans)
Peach Stones/Pits [12]
The peach pit has a composition of biological polymers like cellulose, lignin, and hemicellulose. These compounds increase the intensity of the hydroxyl and phenol groups which give it the adsorbing quality. Additionally, it has a large surface area for binding sites. Modified peach pit was shown to have a better binding affinity for Zearalenone compared to unmodified peach pit.
Charcoal and clay binders have been shown to bind ZON, but studies were done in vitro:
"This study was conducted to evaluate the ability (adsorptive power) of five adsorbents--activated carbon, bentonite, talc, sandstone, and calcium sulfate--to trap Zearalenone in vitro. Activated carbon was the best adsorbent, binding 100% Zearalenone (pH 3 and 7.3) at 0.1, 0.25, 0.5, and 1% dose levels. Bentonite, talc,and calcium sulfate were less efficient than activated carbon but still could bind Zearalenone to some extent."[7]
Bacteria (L. plantarum, Bacillus) [10,11,14,16,17,18]
B. lincheniformis, B. megaterium, B. cereus - BC7, B. subtillus, and B. thuringiensis) were efficacious in binding Zearalenone with the addition of manganese.
Lactic acid bacteria with high esterase activity may be helpful (e.g. Lactobacillus plantarum, specifically Lactobacillus plantarum BCC 47723 A)
Detoxification Pathways for Zearalenone
Zearalenone primarily uses the glucuronidation, sulfation, and methylation Phase II detox pathways. Within the glucuronidation pathway, Zearalenone uses the UGT1A1, UGT1A3, UGT1A8 genes for detoxification. [100]Therefore, understanding your genetic status for key genes in these pathways is helpful. Avoiding foods and compounds that inhibit these pathways will also be helpful, as is paying attention to your nutritional status for the key cofactors in these pathways. Lastly, designing a diet and supplement regimen to help activate or up-regulate these pathways is important.
Oxidative Stress in ZON Toxicity
"Our results demonstrated that Zearalenone inhibits cell proliferation which was accompanied by an increase in the generation of free radicals as measured by fluorescent 2,7-dichlorofluorescein (DCF) and Malondialdehyde (MDA). As an adaptive response to this redox status, we showed an induction of heat shock protein expression (Hsp 70) and an activation of antioxidant enzymes; catalase and Superoxide Dismutase (SOD). Moreover, a loss of mitochondrial membrane potential (Δѱm) was observed."[1]
Estrogen Related Imbalances Secondary to ZON Exposure May Lead to Reproductive System Diseases
"One of these mycotoxins is zearalenone, which is classified as a xenoestrogen, an exogenous compound which resembles the structure of naturally occurring estrogens with its chemical structure. This property of zearalenone determines its ability to bind to estrogen receptors of cell and its bioaccumulation. This leads to disorders of the hormonal balance of the body, which in consequence may lead to numerous diseases of reproductive system such as prostate, ovarian, cervical or breast cancers. High risk posed by long-term exposure to contaminated food forces the modern science to develop and implement effective methods of zearalenone neutralisation."[13]
Zearalenone Induced Toxicity, Oxidative Stress, and Apoptosis In Human Embryonic Stem Cells
"Moreover, differentiation of human embryonic stem cell (hESC) was initiated by embryoid bodies (EBs) formation; EBs were exposed to different concentrations of Zearalenone for 24 h to detect cellular reactive oxygen species (ROS) generation, the mitochondrial transmembrane potential (MMP), apoptosis, cell cycle, and the related protein expression. Based on the results of the three endpoints and functions of Zearalenone in mEST and hEST, Zearalenone was evaluated to have strong embryonic toxicity both by two models. The increases in cellular ROS and loss of MMP were observed at 2 and 4 μg/ml concentrations. Flow cytometry showed that Zearalenone induced cell cycle arrest and apoptosis. The upregulation of p53, caspase-9, caspase-3, and the ratio of Bax/Bcl-2 were observed at 2 and 4 μg/ml concentrations. Collectively these results demonstrate that Zearalenone has strong embryonic toxicity and induces oxidative stress and apoptosis in differentiated human ESCs." [20]
Oxidative Stress and Ferroptosis
Recently, it has been reported that mycotoxins such as deoxynivalenol and zearalenone can induce ferroptosis in the reproductive organs by reducing antioxidant capacity, down-regulating the expression levels of glutathione peroxidase 4 (GPX4) and ferritin heavy chain 1 (FTH1), disrupting iron homeostasis and accumulating lipid peroxidation, ultimately resulting in spermatogenesis damage. [15]
Zearalenone disrupts Copper Transporters ATP7A, ATP7B
"The expression levels of ATPase copper transporting alpha (ATP7A) and ATPase copper transporting beta (ATP7B) were significantly downregulated (p < 0.01), while the expression of solute carrier family 31 member 1 (SLC31A1) was not modified in the ZEA group compared with the NC group." [8]
Potential Supplement Strategies
Antioxidants:
NAC (N-acetyl cysteine). NAC, for most people, is the limiting compound for the creation of glutathione, the body’s master antioxidant
"Porcine small intestinal epithelial (SIEC02) cells were selected to assess the effect of Zearalenone exposure on the intestine. Cells were exposed to Zearalenone (20 µg/mL) or pretreated with (81, 162, and 324 µg/mL) N-acetylcysteine (NAC) prior to Zearalenone treatment. Results indicated that the activities of glutathione peroxidase (Gpx) and glutathione reductase (GR) were reduced by Zearalenone, which induced reactive oxygen species (ROS) and malondialdehyde (MDA) production. Moreover, these activities increased apoptosis and mitochondrial membrane potential (ΔΨm), and regulated the messenger RNA (mRNA) expression of Bax, Bcl-2, caspase-3, caspase-9, and cytochrome c (cyto c). Additionally, NAC pretreatment reduced the oxidative damage and inhibited the apoptosis induced by Zearalenone. It can be concluded that Zearalenone-induced oxidative stress and damage may further induce mitochondrial apoptosis, and pretreatment of NAC can degrade this damage to some extent."[5]
Natural Antioxidants
Shown to modulate oxidative stress and have some benefit with Zearalenone are vitamins A, C, and E, as well as the compounds curcumin and lycopene. [6]
Selenium:
Selenium has been shown to prevent Zearalenone damage to chicken spleen lymphocytes. Treatment with selenium blocked ROS generation, improved anti oxidative capacity, and reversed apoptosis and estrogen receptor stress-related genes and protein expression. Most glutathione enzymes use selenium as their cofactor
"The results show that Zearalenone induced an increase in ROS generation and lipid peroxidation, and a decrease in levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione (GSH). The results of apoptosis morphologically from acridine orange/ethidium bromide (AO/EB) fluorescent staining and flow cytometry analysis show apparent apoptosis in the Zearalenone-treated group, and was confirmed by the upregulation of caspase-3, -12 and downregulation of Bcl-2. Meanwhile, Zearalenone activated the endoplasmic reticulum (ER) stress by upregulating ER stress-related molecular sensors (GRP78, ATF6, ATF4, IRE). However, co-treatment with Se effectively blocked ROS generation, improved anti oxidative capacity, and reversed apoptosis and ER stress-related genes and protein expression. Taken together, these data suggest that oxidative stress and ER stress play a vital role in Zearalenone-induced apoptosis, and Se had a significant preventive effect on Zearalenone-induced apoptosis in chicken spleen lymphocyte via ameliorating the ER stress signaling pathway."[4]
Silymarin / Milk Thistle
Silymarin is a group of constituents from the milk thistle plant, Silybum marinanum
In a rat study, Silymarin has been shown to be protective in the face of Zearalenone-induced liver toxicity and reproductive toxicity. This might be due to the herb’s ability to improve antioxidant capacity and regulate genes related to protein synthesis, Zearalenone metabolism, hormone synthesis, and ABC transporters in the tissues. Helps keep the Bile Salt Export Pump (BSEP) open to avoid 'herx' reactions from high estrogen or overloaded MRP2 proteins that escort toxins into the bile.
"Dietary silymarin supplementation at 100, 200, and 500 mg/kg protected rats from Zearalenone-induced hepatotoxicity and reproductive toxicity, potentially through improvement in the antioxidant capacity and regulation in the genes related to protein synthesis, Zearalenone metabolism, hormone synthesis, and ABC transporters in the tissues" [3]
Melatonin
Melatonin reduces oxidative stress in bovine ovarian granulosa cells
"This study investigated the effects of β-zol and HT-2 on bovine ovarian granulosa cells (BGCs), and how melatonin may counteract these effects. β-zol and HT-2 inhibited cell proliferation in a dose-dependent manner, and induced apoptosis of BGCs. They also yielded upregulation of the apoptosis-related genes Bax/Bcl-2 and Caspase3 and phosphorylation of p38MAPK. Increases in intracellular ROS were observed along with higher levels of mRNA anti-oxidation markers SOD1, SOD2, and CAT. SOD1, SOD2, malonaldehyde (MDA), and glutathione peroxidase (GSH-px) activities increased, as did the levels of SOD1 and SOD2 proteins. All of these effects were reduced or entirely attenuated in BGCs pre-treated with melatonin. Our results demonstrate that melatonin has protective effects against mycotoxin-induced apoptosis and oxidative stress in BGCs"[2]
Chrysin [6]
Chrysin is a flavonoid found in many plants. Chrysin is found in honey, propolis and passion flower.
In one study, chrysin was given to mice at the same time they were given Zearalenone. Chrysin attenuated the toxic effects caused by Zearalenone in blood and testes of mice. Chrysin treatment increased the number and motility of sperm, testosterone levels, restored antioxidant defenses and reduced the inflammation and apoptosis process.
Kefir
Kefir is a fermented drink that contains live bacteria and yeast
"The co-treatment as well as the pre-treatment by kefir showed a reduction of Zearalenone induced damages for all tested markers. However, the pre-treatment seems to be the most efficient, it prevented almost all Zearalenone hazards. Consequently, oxidative damage appears to be a key determinant of Zearalenone induced toxicity in cultured HCT-116 cells. In conclusion, we showed that kefir may better exert its virtue on preventive mode rather than on curative one." [1]
Kefir will usually exacerbate histamine issues if they are present; proceed with caution.
DIM (Diindolymethane)
DIM is a compound found in cruciferous vegetables that may inhibit the conversion of harmful estrogen (16alpha-hydroxyestrone) into a beneficial form (2-hydroxyestrone). High estrogen is implicated in the inhibition of the Bile Salt Export Pump BSEP - (ABCB11), which is controlled by the FXR Receptor (NR1H4), resulting in 'herx' reactions.
If you would like to review your genetics, look at testing options, or discuss strategies to expedite the clearance of Zearalenone, please contact me to set up an appointment. I encourage you to look into the references cited below should you feel called to explore this topic in more detail.
References:
Protective effects of kefir against zearalenone toxicity mediated by oxidative stress in cultured HCT-116 cells. Emna El Golli-Bennour, Rim Timoumi, et. al. Toxicon. 2019 Jan:157:25-34.
Melatonin alleviates β-zearalenol and HT-2 toxin-induced apoptosis and oxidative stress in bovine ovarian granulosa cells. Fangxiao Yang 1, Lian Li, et. al. Environ Toxicol Pharmacol. . 2019 May:68:52-60. doi: 10.1016/j.etap.2019.03.005. Epub 2019 Mar 7.
The Journal of Nutrition Biochemical, Molecular, and Genetic Mechanisms Dietary Silymarin Supplementation Alleviates Zearalenone-Induced Hepatotoxicity and Reproductive Toxicity in Rats Xin Gao, Zhuo-Hui Xiao. 2018 American Society for Nutrition. 1209 First published online August 3, 2018; doi: https://doi.org/10.1093/jn/nxy114
Protective effects of selenium against zearalenone-induced apoptosis in chicken spleen lymphocyte via an endoplasmic reticulum stress signaling pathway. Xiau, Xu, et al. Published: 29 October 2018. Cell and Stress Chaperones. Pages 77–89, Articles/PMC6215273/.
Protective Effect of N-Acetylcysteine against Oxidative Stress Induced by Zearalenone via Mitochondrial Apoptosis Pathway in SIEC02 Cells. . Wang, Li, et. al. Toxins (Basel). 2018 Oct; 10(10): 407. Published online 2018 Oct 9. doi: 10.3390/toxins10100407
Mycotoxins and oxidative stress: where are we? E.O. da Silva1, A.P.F.L. Bracarense1* and I.P. Oswald. World Mycotoxin Journal, 2018; 11 (1): 113-133 Wageningen Academic Publishers SPECIAL ISSUE: 10 years World Mycotoxin Journal ISSN 1875-0710 print, ISSN 1875-0796 online, DOI 10.3920/WMJ2017.2267 113 1.
In vitro binding of zearalenone to different adsorbents. Dante J Bueno 1, Liliana Di Marco. J Food Prot. . 2005 Mar;68(3):613-5. doi: 10.4315/0362-028x-68.3.613. PMID: 15771192. DOI: 10.4315/0362-028x-68.3.613
Effects of Zearalenone on Apoptosis and Copper Accumulation of Goat Granulosa Cells In Vitro. Lie, Ma, et. al. Biology (Basel). 2023 Jan; 12(1): 100. Published online 2023 Jan 9. doi: 10.3390/biology12010100. PMCID: PMC9856194. PMID: 36671791
Zearalenone, an estrogenic mycotoxin, is an immunotoxic compound
Isolation and characterization of the Bacillus cereus BC7 strain, which is capable of zearalenone removal and intestinal flora modulation in mice. Yue Wang , Jian Zhang, et al.. Toxicon. . 2018 Dec 1:155:9-20. doi: 10.1016/j.toxicon.2018.09.005. Epub 2018 Sep 26.
Characterization of a cold-active esterase from Lactobacillus plantarum suitable for food fermentations. María Esteban-Torres 1, José Miguel Mancheño, et. al. J Agric Food Chem.
IN VITRO EVALUATION OF THE EFFICACY OF PEACH STONES AS MYCOTOXIN BINDERS. Z o r i c a R. L o p i č i ć 1 , A l e k s a n d r a S. B o č a r o v – S t a n č i ć 2 , M i r j a n a D. S t o j a n o v i ć, et. al. Jour. Nat. Sci, Matica Srpska Novi S № 124, 287—296, 2013. UDC 634.25:544.725 DOI: 10.2298/ZMSPN1324287L.
Zearalenone and its metabolites: Effect on human health, metabolism and neutralisation methods. A Rogowska 1, P Pomastowski. Toxicon . 2019 Apr 15:162:46-56. doi: 10.1016/j.toxicon.2019.03.004. Epub 2019 Mar 6. PMID: 30851274. DOI: 10.1016/j.toxicon.2019.03.004
14. Qin, X., Su, X., Tu, T., Zhang, J., Wang, X., Wang, Y., Wang, Y., Bai, Y., Yao, B., Luo, H., & Huang, H. (2021). Enzymatic degradation of multiple major mycotoxins by dye-decolorizing peroxidase from bacillus subtilis. Toxins, 13(6), 429. https://doi.org/10.3390/toxins13060429
15. Effect of Zearalenone-Induced Ferroptosis on Mice Spermatogenesis. Li, Zhu, et al. Animals (Basel) . 2022 Nov 3;12(21):3026. PMID: 36359150. PMCID: PMC9657494. DOI: 10.3390/ani12213026.
16. Detoxification Strategies for Zearalenone Using Microorganisms: A Review. Wang, Wu, et. al. Microorganisms. 2019 Jul; 7(7): 208. Published online 2019 Jul 21. doi: 10.3390/microorganisms7070208. PMCID: PMC6680894. PMID: 31330922
17. Huang,W.; Chang, J.;Wang, P.; Liu, C.; Yin, Q.; Zhu, Q.; Lu, F.; Gao, T. Effect of the combined compound probiotics with mycotoxin-degradation enzyme on detoxifying aflatoxin B1 and zearalenone. J. Toxicol. Sci. 2018, 43, 377–385. Microorganisms 2019, 7, 208 14 of 14
18. Huang, W.; Chang, J.; Wang, P.; Liu, C.; Yin, Q.; Song, A.; Gao, T.; Dang, X.; Lu, F. Effect of compound probiotics and mycotoxin degradation enzymes on alleviating cytotoxicity of swine jejunal epithelial cells induced by aflatoxin B1 and zearalenone. Toxins (Basel) 2019, 11, 12.
19. Zearalenone induces NLRP3-dependent pyroptosis viaactivation of NF-κB modulated by autophagy in INS-1 cells. Xue Wang, Liping Jiang ,Toxicology. Volume 428. 1 December 2019 , 152304. Science Direct. https://doi.org/10.1016/j.tox.2019.152304
20. Zearalenone causes embryotoxicity and induces oxidative stress and apoptosis in differentiated human embryonic stem cells. Hanwen Cao , Yuan Zhi, et. al. Toxicology in Vitro. Volume 54, February 2019, Pages 243-250. https://doi.org/10.1016/j.tiv.2018.09.020
21. Effects and Underlying Mechanisms of Zearalenone Mycotoxin at Concentrations Close to the EC Recommendation on the Colon of Piglets after Weaning, by Valeria Cristina Bulgaru
22. Glucuronidation of zearalenone, zeranol and four metabolites in vitro: formation of glucuronides by various microsomes and human UDP-glucuronosyltransferase isoforms Erika Pfeiffer 1, Andreas Hildebrand, et. al. Mol Nutr Food Res. . 2010 Oct;54(10):1468-76. doi: 10.1002/mnfr.200900524. PMID: 20397195. DOI: 10.1002/mnfr.200900524
23. Exposure to Zearalenone Leads to Metabolic Disruption and Changes in Circulating Adipokines Concentrations in Pigs. Toxins (Basel). 2021 Nov; 13(11): 790. Published online 2021 Nov 8. doi: 10.3390/toxins13110790 PMCID: PMC8618343 PMID: 34822574